Abstract
Acetoin is widely used in food and other industries. A bdhA and acoA double-knockout strain of Bacillus subtilis produced acetoin at 0.72 mol/mol, a 16.4 % increased compared to the wild type. Subsequent overexpression of the alsSD operon enhanced the acetolactate synthase activity by 52 and 66 % in growth and stationary phases, respectively. However, deletion of pta gene caused little increase of acetoin production. For acetoin production by the final engineered strain, BSUW06, acetoin productivity was improved from 0.087 g/l h, using M9 medium plus 30 g glucose/l under micro-aerobic conditions, to 0.273 g/h l using LB medium plus 50 g glucose/l under aerobic conditions. In fermentor culture, BSUW06 produced acetoin up to 20 g/l.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Fradrich C, March A, Fiege K, Hartmann A, Jahn D, Hartig E (2011) The transcription factor AlsR binds and regulates the promoter of the alsSD operon responsible for acetoin formation in Bacillus subtilis. J Bacteriol 194:1110–1112
Holtzclaw WD, Chapman LF (1975) Degradative acetolactate synthase of Bacillus subtilis: purification and properties. J Bacteriol 121:917–922
Huang M, Oppermann-Sanio FB, Steinbuchel A (1999) Biochemical and molecular characterization of the Bacillus subtilis acetoin catabolic pathway. J Bacteriol 181:3837–3841
Ji XJ, Huang H, Li S, Du J, Lian M (2007) Enhanced 2,3-butanediol production by altering the mixed acid fermentation pathway in Klebsiella oxytoca. Biotechnol Lett 30:731–734
Liu YF, Zhang SL, Yong YC, Ji ZX, Ma X, Xu ZH, Chen SW (2011) Efficient production of acetoin by the newly isolated Bacillus licheniformis strain MEL09. Process Biochem 46:390–394
Moes J, Griot M, Keller J, Heinzle E, Dunn IJ, Bourne JR (1984) A microbial culture with oxygen-sensitive product distribution as a potential tool for characterizing bioreactor oxygen transport. Biotechnol Bioeng 27:482–489
Nicholson WL (2008) The Bacillus subtilis ydjL (bdhA) gene encodes acetoin reductase/2,3-butanediol dehydrogenase. Appl Environ Microbiol 74:6832–6838
Renna MC, Najimudin N, Winik LR, Zahler SA (1993) Regulation of the Bacillus subtilis alsS, alsD, and alsR genes involved in post-exponential-phase production of acetoin. J Bacteriol 175:3863–3875
Silbersack J, Jürgen B, Hecker M, Schneidinger B, Schmuck R, Schweder T (2006) An acetoin-regulated expression system of Bacillus subtilis. Appl Microbiol Biotechnol 73:895–903
Thanh TN, Jürgen B, Bauch M, Liebeke M, Lalk M, Ehrenreich A, Evers S, Maurer KH, Antelmann H, Ernst F, Homuth G, Hecker M, Schweder T (2010) Regulation of acetoin and 2,3-butanediol utilization in Bacillus licheniformis. Appl Microbiol Biotechnol 87:2227–2235
Werpy T, Petersen G (2004) Top value added chemicals from biomass: Volume I: results of screening for potential candidates from sugars and synthesis gas. US Department of Energy
Xu P, Xiao Z, Du Y, Wei Z (2009) Acetoin high-yield Bacillus pumilus strain. US Patent Application 20090215152
Xu H, Jia S, Liu J (2011) Development of a mutant strain of Bacillus subtilis showing enhanced production of acetoin. Afr J Biotechnol 10:779–788
Yang YT, Peredelchuk M, Bennett GN, San KY (2000) Effect of variation of Klebsiella pneumoniae acetolactate synthase expression on metabolic flux redistribution in Escherichia coli. Biotechnol Bioeng 69:150–159
Zhang L, Yang Y, Sun J, Shen Y, Wei D, Zhu J, Chu J (2010) Microbial production of 2,3-butanediol by a mutagenized strain of Serratia marcescens H30. Bioresour Technol 101:1961–1967
Zhu Y, Chen X, Chen T, Zhao X (2007) Enhancement of riboflavin production by overexpression of acetolactate synthase in a pta mutant of Bacillus subtilis. FEMS Microbiol Lett 266:224–230
Acknowledgments
This work was supported by National Natural Science Foundation of China (NSFC-21176182), Natural Science Foundation of Tianjin (12JCYBJC12900), the Research Fund for the Doctoral Program of Higher Education (20100032120014) and the National 973 Project (2011CBA00804, 2012CB725203).
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Wang, M., Fu, J., Zhang, X. et al. Metabolic engineering of Bacillus subtilis for enhanced production of acetoin. Biotechnol Lett 34, 1877–1885 (2012). https://doi.org/10.1007/s10529-012-0981-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-012-0981-9